Antral Fluid Research Articles

STUDY TO COMPARE FASTING GASTRIC VOLUME USING ULTRASOUND IN DIABETIC AND NON DIABETIC PATIENTS POSTED FOR ELECTIVE SURGERIES

Objective: To study the comparison of fasting gastric volume using ultrasound in diabetic and nondiabetic patients in elective surgeries. To compare the incidence of delayed gastric emptying in diabetic and nondiabetic after standard fasting guidelines Material and Methods: After institutional ethical committee approval, CTRI registration and patient consent, prospective study was conducted in 30 diabetic and 30 non-diabetic patients aged 30-60years of American society of anesthesiologists- physical status I-III having similar fasting intervals. Pre operatively, quantitative assessments of gastric antrum in supine and right lateral decubitus (RLD) positions were performed using curvilinear probe in accordance to standard gastric scanning. USG grade, cross-sectional area (CSA) of the antrum and gastric volume were calculated. The gastric antrum was classified as Grade 0, 1 or 2, signifying empty antrum, fluid in RLD position only and antral fluid in both supine and RLD positions, respectively. Results:The CSA of 2.35 ± 1.07 cm2 and 3.51 ± 1.4 cm2 in diabetic were significantly higher (P = 0.001) than 1.37 ± 0.32 cm2 and 2.19 ± 1.15 cm2 of control, in supine and RLD positions, respectively. GV was 3.85± 20.06 ml in control group and 6.5±23.51 ml in diabetic group. Conclusion: Diabetic patients have higher gastric antral cross-sectional area and gastric volumes as observed by gastric ultrasound than the non-diabetic patients. Reference - 1. Robinson M, Davidson A. Aspiration under anaesthesia: Risk assessment and decision-making. Contin Edu Anaesth Crit Care Pain. 201314:171–5. 2.Perlas A, Chan VW, Lupu CM, Mitsakakis N, Hanbidge A. Ultrasound assessment of gastric content and volume. Anesthesiology. 2009111:82–9. 3. Smith I, Kranke P, Murat I, Smith A, OSullivan G, Soreide E, et al. Perioperative fasting in adults and children: Guidelines from the European Society of Anaesthesiology. Eur J Anaesthesiol. 201128:556–69.

ACTIVITY AND CONTENT OF SUPEROXIDEDISMUTASE IZOZYMES IN GRANULOSE CELLS FROM COW OVARY FOLLICLES

The activity and content of superoxide dismutase isoforms (SOD) in granulosa cells from cow ovarian follicles were studied for research after slaughter of cows ovaries were selected, which according to the physiological state were divided into groups: with "fresh" ovulation, at the site of the ovulated follicle there is a hole, no corpus luteum or diameter up to 5 mm, red color (n = 14); with "early corpus luteum", diameter 10-20 mm, color red or brown (n = 41); with “late corpus luteum", diameter 5–15 mm, color yellow (n = 32); "follicular growth", without the corpus luteum (n = 84). The ovaries of cows with small (<4 mm), medium (4 - 7 mm) and large (> 7 mm) follicles were used. Antral fluid was obtained from the follicles, from which granulosa cells were isolated. Cells were suspended according to the volume of follicular fluid in the medium Dulbeccos modified Eagle medium (DME) with the addition of estrus serum of cows, follicular fluid, insulin and heparin. In cell culture, protein concentration, activity, and superoxide dismutase isozymes were determined. It was found that granulosa cells are characterized by SOD activity - 12.4 ± 0.74 IU / mg protein (6.8 ± 1.72 - 19.8 ± 3.75 IU / mg protein). The activity of SOD in the culture of granulosa cells had 5–6 isoforms of the enzyme. It was found that isoforms at the site of localization are divided into cytosolic, mitochondrial and extracellular proteins of SOD. The cytosolic isoform were represented by 3 - 4, and mitochondrial and extracellular have one active protein of the enzyme. he activity of the enzyme and the content of isoforms depended on the size of the follicles from which the cells are removed and the physiological state of the ovaries. The studied indicators characterize the intensity of oxidative metabolism as a whole in cells and in their individual parts and organelles. For cultivation, it is advisable to select granulosa cells from ovarian follicles of "follicular growth" and "late corpus luteum" because they are characterized by consistently high activity of SOD, which protects intracellular components from the cytotoxic action of superoxide anion.

Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats.

ObjectiveTyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited.MethodsThe ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blot analysis, respectively.ResultsTyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins.ConclusionGiven that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

Intra‐follicular temperature acts to regulate mammalian ovulation

Writing as one with a long-standing interest in the process of mammalian ovulation,1, 2 any new evidence as to factors underlying the release of an oocyte from a mature Graafian follicle is welcome. We now wish to report that one such factor is temperature of the antral fluid, and that this temperature corresponds closely with, or mirrors, that of the ovarian follicular surface. There is a cumulative body of evidence that pre-ovulatory follicles may be more than 1.0°C cooler than ovarian stroma in eutherian mammals, and both compartments are cooler than jugular and deep rectal temperatures.3, 4 Greater confidence in the work and its potential significance has now been achieved in mature cows.5-7 Our recent findings can be summarized in a single sentence. Follicles having a reduced antral fluid temperature compared with that of neighboring tissues invariably ovulated whereas follicles not revealing such cooling did not. This evidence comes from cow ovaries which, it should be noted, have various features in common with human ovaries: these include the pre-ovulatory follicular diameter (commonly 14-22 mm) and the time interval from the gonadotropin peak to ovulation itself (approximately 26-36 hours). Previous studies in rabbits (n = 40),8 pigs (n = 89),1, 9 cows (n = 6)10 and women (n = 26)11 employed infra-red sensing of surface temperatures8-10, 12 or follicular puncture with thermoelectrodes11 in ovaries exposed during mid-ventral or flank laparotomy under systemic or local anesthesia. All follicles in the different studies except one (from an infertile woman)11 were cooler than the ovarian stroma. In a series of recent studies, by contrast, a fine thermistor probe was introduced into the follicular antrum of dairy cows (n = 80) without the use of sedatives, anesthetics or surgery.5-7 The only tissue invasion involved passing a 17G needle containing the probe across the anterior vaginal wall followed by puncture of the ovarian surface. The procedure was monitored under ultrasonic scanning. None of the animals revealed any form of discomfort or stress during or after this straightforward approach. Results are summarized in Table 1. Graafian follicles that ovulated within 24 hours of temperature measurement were a mean of 1.12°C cooler than deep rectal temperatures, whereas no significant differences were found for non-ovulating follicles. This lower temperature was a consistent observation throughout the studies,5-7 enabling follicular cooling to be correlated positively with the potential for pregnancy.7 Evidence in support of this point can be detected in infra-red images of pig ovaries.12 By contrast, the temperature of newly ovulated follicles—ie, without their content of cumulus-oocyte complex and follicular fluid—was similar to that of other ovarian tissues. Pregnancies n (%) The specific mechanisms involved in follicular cooling have not been fully revealed, although they currently focus on endothermic reactions within pre-ovulatory follicles and would require effective local heat exchange.13, 14 The present results should stimulate further examination of antral fluid contents, especially large glycoprotein molecules, during the remodeling that leads to increased viscosity. At a more practical level, these results provide much-needed inspiration for human in vitro fertilization clinics because they offer a meaningful route to oocyte selection in protocols of multiple follicle stimulation by systemic gonadotropin treatment.15 Of course, non-invasive infra-red scanning of ovarian surface temperature would be a more attractive approach than multiple follicle puncture, but this awaits an endoscope capable of transmitting infra-red radiation. Relationships between temperature, follicular growth in vitro and oocyte quality would also be considered in the procedures of fertility preservation.16-19

Comparison of fasting gastric volume using ultrasound in diabetic and non-diabetic patients in elective surgery: An observational study.

Background and Aims:Gastroparesis despite standard fasting in diabetic patients may increase the aspiration risk. This study aimed to compare fasting gastric volume (GV) of diabetic with non-diabetic patients scheduled for elective surgery using USG.Methods:This prospective observational study included 53 diabetic and 50 non-diabetic patients aged >18 years, American Society of Anesthesiologists' physical status I-III having similar fasting intervals. Before induction, using standard gastric scanning protocol, qualitative and quantitative assessments of gastric antrum in supine and right lateral decubitus (RLD) positions were performed with a curved array probe. USG grade, cross-sectional area (CSA) of the antrum and GV were calculated. The gastric antrum was classified as Grade 0, 1 or 2, signifying empty antrum, fluid in RLD position only and antral fluid in both supine and RLD positions, respectively.Results:In supine position, CC and AP diameters were 1.96 ± 0.41 cm and 0.9 ± 0.57 cm in control group and 2.28 ± 0.50 cm and 1.39 ± 0.44 cm in diabetic group, respectively. In RLD, CC was 2.28 ± 0.57 cm and AP was 1.24 ± 0.42 cm in control group as compared to CC 2.54 ± 0.56 cm and AP 1.82 ± 0.56 cm in diabetic group. The CSA of 2.57 ± 1.19 cm2 and 3.73 ± 1.61 cm2 in diabetic were significantly higher (P = 0.001) than 1.41 ± 0.55 cm2 and 2.30 ± 1.18 cm2 of control, in supine and RLD positions, respectively. GV was 4.20 ± 22.26 ml in control group and 9.15 ± 25.70 ml in diabetic group.Conclusion:Diabetic patients have higher gastric antral cross-sectional area and gastric volumes as observed by gastric ultrasound than the non-diabetic patients.

Biochemical constituents and steroid hormones in follicular fluid from antral follicles of cross-bred Malabari goats of Kerala

ABSTRACT A study was conducted in cross-bred (CB) Malabari goats of Southern India, Kerala to assess the pattern of change in biochemical constituents of follicular fluid during final stages of follicle development. One hundred and twenty goat ovaries were collected from local slaughter houses. After classifying the antral follicles based on size i.e. small (1–3 mm) and large (3–6 mm), the follicular fluid was aspirated and analysed to estimate the concentration of various biochemical constituents including total protein, albumin, globulin, glucose, triglycerides, cholesterol, high-density lipoprotein (HDL) and steroid hormones. The results showed that the concentration of glucose in follicular fluid was significantly higher (p < 0.05) in small follicles compared the large ones. However, the level of HDL and oestrogen was significantly higher (p < 0.05) in large follicles. Follicular fluid protein, albumin, globulin, triglycerides, cholesterol, and progesterone did not differ significantly between the small and large follicles. The results of the present study suggest that the biochemical constituents in the antral follicular fluid vary with the follicular size and the data would contribute to better understanding of the follicular fluid metabolite dynamics of goat ovarian follicles which imparts oocyte competence for fertilization during later developmental stages.

Proposed Clinical Decision Rules to Diagnose Acute Rhinosinusitis Among Adults in Primary Care.

To reduce inappropriate antibiotic prescribing, we sought to develop a clinical decision rule for the diagnosis of acute rhinosinusitis and acute bacterial rhinosinusitis. Multivariate analysis and classification and regression tree (CART) analysis were used to develop clinical decision rules for the diagnosis of acute rhinosinusitis, defined using 3 different reference standards (purulent antral puncture fluid or abnormal finding on a computed tomographic (CT) scan; for acute bacterial rhinosinusitis, we used a positive bacterial culture of antral fluid). Signs, symptoms, C-reactive protein (CRP), and reference standard tests were prospectively recorded in 175 Danish patients aged 18 to 65 years seeking care for suspected acute rhinosinusitis. For each reference standard, we developed 2 clinical decision rules: a point score based on a logistic regression model and an algorithm based on a CART model. We identified low-, moderate-, and high-risk groups for acute rhinosinusitis or acute bacterial rhinosinusitis for each clinical decision rule. The point scores each had between 5 and 6 predictors, and an area under the receiver operating characteristic curve (AUROCC) between 0.721 and 0.767. For positive bacterial culture as the reference standard, low-, moderate-, and high-risk groups had a 16%, 49%, and 73% likelihood of acute bacterial rhinosinusitis, respectively. CART models had an AUROCC ranging from 0.783 to 0.827. For positive bacterial culture as the reference standard, low-, moderate-, and high-risk groups had a likelihood of acute bacterial rhinosinusitis of 6%, 31%, and 59% respectively. We have developed a series of clinical decision rules integrating signs, symptoms, and CRP to diagnose acute rhinosinusitis and acute bacterial rhinosinusitis with good accuracy. They now require prospective validation and an assessment of their effect on clinical and process outcomes.

Активність та вміст ізозимів аспартатамінотрансферази в репродуктивних органах корів

Activity and isozyme content of aspartate transaminase in cow reproductive organs at different physiological states and ovarian hypofunction were studied. It was registered, that activity of aspartate transaminase (AST) in ovarian tissue and endometrium at corpus luteum state is 132.0 ± 8.23 and 172.9 ± 8.70 μm/min×mg of protein, lower on 26.3 and 5.5% at follicle growth and at early corpus luteum is 66.5 ± 2.10 and 147.1 ± 13.26 μm/min×mg of protein. Lowest AST activity is registered at hypofunction state (61.8 ± 4.57 і 146.1 ± 9.33 μm/min×mg of protein). In cow reproductive organ tissues and antral fluid from ovarian follicles was registered two bands of proteins, that have AST activity. In reproductive organs main part is mitochondrial isozyme (АSТ2; 78.8–97.7%) in antral fluid – cytosole (АSТ1; 72.8–88.2%). In ovarian and uterus tissues was established high enzyme activity at late corpus luteum, that characterizes intensive aspartate conversion to oxaloacetate and possible utilization of substeates to maintain energetic and sytnthetic needs in cow reproductive organs. Antral fluid of big follicles from ovarian at follicle growth and at state of late corpus luteum characterizes by high activity and content of AST, that points on follicle cell growth – granulose and oocyte. In ovarian tissue and endometrium at hypofunction lower activity and isozyme content point on substrate deficiency for transamination and violation of physiological functions.

Relationship between endometritis and oxidative stress in the follicular fluid and luteal function in the buffalo.

In this study, alteration in the follicular fluid composition and luteal function was investigated in the buffalo with endometritis. Genitalia were classified into cytological and purulent endometritis on the basis of polymorphonuclear cell cut off while non-endometritis served as control (n=10/group). In the follicular phase, the number of surface follicles was counted, diameter of the largest follicle was measured and the follicular fluid was assayed for total protein, cholesterol, malondialdehyde (MDA), total antioxidant capacity (TAC), oestradiol (E2 ) and progesterone (P4 ). The P4 content of corpus luteum during mid-luteal phase was estimated by radioimmunoassay. Ovaries from the follicular phase of oestrous cycle showed no significant difference in the total number of surface follicles, size of the largest follicle and volume of follicular fluid in the buffaloes with and without endometritis (p>.05). However, the antral fluid of the largest follicle from the genitalia of buffalo with cytological and purulent endometritis showed a significant decrease in the concentration of total protein, cholesterol, TAC and E2 and a significant increase in the concentration of MDA and P4 (p<.05). The results indicated that there is an association between endometritis and decreased ovarian function.

Ovarian Antral Follicular Dynamics and Follicular Fluid Composition in Non-descript Goats of Karnataka

A study was carried out to elucidate the ovarian antral follicular dynamics and the profile of certain biochemical components in ovarian antral follicular fluid of non-descript goats of Karnataka. Three hundred ovaries were collected from two to five year old healthy non-descript goats slaughtered at Civil Meat Processing and Production Centre, Bangalore during the months of September to December, 2012. The surface ovarian antral follicles were categorized into three groups based on the diameter as Group I (small, 1 to 2.9 mm), Group II (medium, 3 to 5 mm) and Group III (large, >5 mm) follicles. The follicular fluid was collected and pooled as per the different groups of follicles and it was analysed for the levels of glucose, total protein, albumin, total cholesterol, estradiol-17β, alkaline phosphatase and acid phosphatase activities. The results of the present study revealed that the number of medium sized follicles were significantly (P 0.05) between various groups of follicles. It was concluded that the findings of the present study on ovarian antral follicular dynamics and biochemical profile of follicular fluid could be useful for the scientists working on in vitro maturation and in vitro fertilization in goats.

Novel follicular-phase gonadotropin-releasing hormone antagonist stimulation protocol for in vitro fertilization in the poor responder

Poor responders continue to be vexing in infertility therapy. By using GnRH antagonists before ovarian stimulation, we demonstrate an improvement in oocyte, embryo, and zygote yield in patients with a prior poor response.

Continuous Combined Letrozole-FSH Stimulation Requires Less FSH With Similar Outcomes Compared To Standard Ovarian Stimulation Regimens for IVF

We have previously described a combined letrozole-FSH stimulation protocol (letFSH) for breast cancer patients undergoing IVF for embryo cryopreservation. Our current objective was to compare this protocol to standard IVF protocols. Prospective study with age matched controls. Twenty-five breast cancer patients were treated by letFSH protocol in which letrozole 5 mg was started on cycle day 2 and FSH 150 IU was added on cycle day 4. Both medications continued until the day of hCG. GnRH antagonist was administered when E2>250pg/mL. Standard protocol group consisted of leuprolide suppression followed by gonadotropins (n=68) or gonadotropins with GnRH antagonists (n=27). Day-3 FSH levels were similar between the letFSH and the age-matched standard IVF protocol patients (table). Despite the significantly lower peak estradiol levels in the letFSH group, endometrial thickness, percent mature oocytes, fertilization rates and number of 2-pronuclei embryos were similar to standard-IVF patients (table). However, hCG had to be administered at a significantly larger follicle diameter to achieve these results. The loess fit analysis showed that an average diameter of approximately 20 mm of the two leading follicles resulted in the best fertilization rates with letFSH. The length of stimulation was not different between letFSH and standard IVF stimulation cycles (table). While letFSH resulted in higher number of ≥17 follicles, 46 % less FSH was required to achieve these results compared to all standard-IVF cycles. When this was compared to the results of the stimulation with gonadotropins+GnRH antagonist, a protocol more comparable to the stimulation in letFSH, the FSH requirement was reduced further by 65%. Tabled 1 Continuous administration of letrozole with FSH results in similar fertilization rates, number of two pronuclear embryos per cycle, and endometrial thickness while significantly reducing the requirement for gonadotropin administration compared to standard ovarian stimulation protocols. The hCG administration criteria has to be modified but since length of stimulation with letFSH is similar to that of with standard IVF, this is likely to be due to altered antral fluid dynamics. In addition to its use for embryo cryopreservation in breast cancer patients prior to chemotherapy, this combined continous letrozole-FSH protocol may be a cost saving alternative to standard ovarian stimulation protocols.

Tumor necrosis factor alpha regulates collagenolytic activity in preovulatory ovine follicles: relationship to cytokine secretion by the oocyte-cumulus cell complex.

The pleiotropic cytokine tumor necrosis factor (TNF)-alpha has been implicated in the mechanism of ovulation. Experiments were designed to test the hypothesis that TNF-alpha secreted from the oocyte-cumulus cell complex stimulates follicular collagenase production and thereby contributes to ovarian wall degradation and ovulatory rupture. Proestrous ewes were treated with GnRH to synchronize the onset of the gonadotropin surge; ovulation occurs approximately 24 h later. There was an increase in TNF-alpha (immunoassay) in antral fluid of preovulatory follicles at 18 h after GnRH, which was related to tissue collagenolytic bioactivity (radiolabeled type I substrate digestion by enzymatic extract) and collagen (hydroxyproline) depletion. Intrafollicular injection of TNF-alpha antibodies at 12 h after GnRH negated the rise in follicular collagenolytic bioactivity (and is known to block ovulation in the sheep). Moreover, collagenase production was enhanced when follicular tissues (0 h GnRH) were incubated (6 h) with recombinant TNF-alpha; this effect was abolished by the transcriptional inhibitor actinomycin D. Secretion of TNF-alpha by oocyte-cumulus cell complexes isolated from preovulatory follicles simulated the in vivo circumstance. Immunostaining indicated that TNF-alpha was confined mainly to the oocyte before GnRH administration, accumulated in cumulus cells during the mid-to-late preovulatory period, and was expended with the imminent approach of ovulation. To our knowledge, this is the first report specifying that up-regulation of collagenase expression is a target mode of TNF-alpha action in preovulatory follicles. The oocyte-cumulus cell complex is an apparent source of soluble TNF-alpha.

Ultrasonic characterization of ovulatory follicular evacuation and luteal development in heifers.

The characteristics of follicle evacuation during ovulation and the development of the corpus luteum until day 5 (day 0 = ovulation) were studied in seven nulliparous Holstein heifers using real-time ultrasonography. Ovulation was induced and synchronized with a single injection of PGF2 alpha followed in 36 h by GnRH. Continuous scanning and videotaping was performed from apparent stigma formation until antral fluid was no longer detected. The beginning of follicular evacuation (second 0) was defined, retrospectively, after the antral area decreased 10% or more in 1 s. The completion of evacuation was defined as the inability to detect the antrum (the beginning of luteal development, 0 h). Corpora lutea development was monitored at 0, 4 and 20 h, and every 24 h thereafter until day 5. Changes in the maximal cross-sectional area of the antrum, luteal tissue, and central luteal cavities and in the pixel intensity of luteal tissue were determined using a computerized image program. The initial antral fluid evacuation occurred in two patterns that could be readily separated: (1) rapid, means of 58 and 89% evacuation in 1 and 4 s, respectively (four heifers); and (2) slow, means of 17 and 35% in 1 and 4 s, respectively (three heifers). The initial loss that distinguished the two patterns involved about 4 and 20 s for rapid and slow evacuation, respectively. Thereafter, the loss patterns were similar for the two types. The time from the beginning to the completion of evacuation ranged from 6 s to 14.5 min. Mean luteal tissue area increased (P < 0.05) between completion of evacuation (91.2 +/- 6.5 mm2) and day 3 (164.4 +/- 13.7 mm2) and between day 3 and day 4 (263.4 +/- 26.6 mm2). The growth rate of the luteal tissue area between day 3 and day 4 (103.2 +/- 16.0 mm2 day-1) was greater (P < 0.05) than that between day 2 and day 3 (41.9 +/- 12.4 mm2 day-1) and between day 4 and day 5 (49.7 +/- 22.0 mm2 day-1). In contrast to increasing luteal tissue area, mean pixel intensity decreased (P < 0.05) progressively between the completion of evacuation (78.4 +/- 6.3) and day 2 (60.4 +/- 2.5) and did not change significantly thereafter. In conclusion, initial follicular fluid loss during ovulation occurred in two patterns, involving about 4 and 20 s, respectively. The most intensive luteal tissue growth occurred between day 3 and day 4, and the echogenicity of the luteal tissue decreased between day 0 and day 2.

Concentration of tissue inhibitor of metalloproteinases (TIMP)-1 in ovine follicular fluid and serum.

Tissue inhibitor of metalloproteinases (TIMP)-1 mRNA and protein localize within granulosal cells of post-gonadotropin-surge follicles and luteal tissue in ewes. Our objectives were to test the hypotheses that 1) follicular fluid concentration of TIMP-1 increases following a gonadotropin surge induced by LHRH agonist (Exp. 1) and 2) luteal status affects peripheral serum concentration of TIMP-1 (Exp. 2 and 3). In Exp. 1, the concentration of TIMP-1 within antral fluid from post-surge follicles (28.7 +/- 6.65 microg/mL) was greater (P < .02) than from pre-surge follicles (2.37 +/- 2.47 microg/mL). In Exp. 2, serum concentration of TIMP-1 did not differ among d 0 to 6 (1.27 +/- .55 microg/mL) of the estrous cycle or among periods of luteal maintenance (1.29 +/- .06 microg/mL), spontaneous luteal regression (1.19 +/- .09 microg/mL), or luteal development (1.22 +/- .08 microg/mL). However, serum concentration of TIMP-1 was greater ( P < .001) during the period of luteal maintenance (1.14 +/- .04 microg/mL) than during PGF2alpha-induced luteolysis (d 26; .85 +/- .06 microg/mL) and induced luteal absence (d 27 to 33; .95 +/- .05 microg/mL). In Exp. 3, ewes (n = 14) were bled daily from d 1 to 19 (d 0 = estrus) and at 12-min intervals for 6 h on d 3, 10, and 17. Although concentration of TIMP-1 varied considerably within and among ewes, mean concentration of TIMP-1 per ewe per day increased ( P < .05) from d 3 to 17. These data indicate that follicular fluid concentration of TIMP-1 increases following a gonadotropin surge, but the contribution of ovarian derived TIMP-1 to peripheral serum concentration is negligible.

Screening for sinus disease in patients with asthma: A computed tomography-controlled comparison of A-mode ultrasonography and standard radiography

Background: Paranasal sinus disease and bronchial asthma are frequently associated. Patients with asthma often have chronic inflammatory changes of the paranasal mucosa rather than acute bacterial sinusitis. Our aim was to compare the rankings of A-mode ultrasonography and standard radiography as routine screening procedures in the diagnostic workup of these patients. Methods: We compared the evaluation of the maxillary sinuses by A-mode ultrasonography and standard radiographs. Computed tomography served as a gold standard in 19 patients with asthma who had no history of sinus surgery. Results: Computed tomography showed at least some minimal mucosal thickening in any of the paranasal sinuses in 74% and of the maxillary sinuses in 61% of the patients. Compared with the results of computed tomography, plain-view radiography gave a specificity of 86.7% for the maxillary sinuses. Although all cases of severe mucosal thickening were detected, sensitivity for minimal mucosal hyperplasia was low, at 52.2%. In contrast, A-mode ultrasonography demonstrated a sensitivity of 70% but a specificity of only 22%. Conclusions: Even symptom-free patients with asthma show a high prevalence of at least limited mucosal thickening in the paranasal sinuses. Acute sinusitis is not a common finding. A-mode ultrasonography does not allow sufficient evaluation of this mucosal hyperplasia and is therefore not suitable for initial screening in these patients. It may prove helpful as a follow-up in selected patients with known anatomic characteristics, however, especially when antral fluid is involved. For routine screening in patients with asthma, a conventional Waters' view radiograph should be used in conjunction with direct visualization of the ostial-meatal area by fiberoptic or rigid rhinoscopy. This combination provides information about the degree of mucosal hyperplasia as well as mucosal inflammation and secretion.

In situ ligand binding of recombinant human [125I] activin-A and recombinant human [125I]inhibin-A to the adult rat ovary.

Inhibin and activin are hormones produced by ovarian follicles. Specific ovarian cells that bind iodinated recombinant human (rh)-activin-A and rh-inhibin-A were identified by in situ ligand binding. Iodinated rh-molecules were incubated with tissue sections from ovaries, uterus, and oviduct, collected on the mornings of metestrus, diestrus, proestrus, and estrus and the evening of proestrus. Additionally, inhibin/activin subunit mRNA and follistatin mRNA accumulation was examined by in situ hybridization of radiolabeled antisense riboprobes. The cellular site of activin/inhibin binding could thus be colocalized to the cellular site of ligand mRNA and binding protein mRNA production. The association of both [125I]rh-activin-A and [125I] rh-inhibin-A with specific cell types varied across the rat estrous cycle. Nonspecific binding was evaluated by competition with a 1000-fold excess of homologous ligand, and low affinity association of the heterologous ligand was evaluated by competition with a 1000-fold excess of heterologous ligand. [125I]rh-activin-A binding was more widespread than was [125I]rh-inhibin-A binding under our experimental conditions. [125I]rh-Activin-A bound to the granulosa cells of all stages of follicles: unrecruited, growing, and Graafian follicles. Thecal cell binding was found in developing follicles (350-500 microns). The granulosa cells of stimulated follicles (evening of proesterus) bound less [125I]rh-activin-A than those of unstimulated follicles. [125I]rh-Activin-A binding was also associated with antral fluid of follicles in each size class. Although early atretic follicles retained some [125I]rh-activin-A binding, late atretic follicles did not bind [125I]rh-activin-A. Corpus luteum present on metestrus and diestrus bound [125I]rh-activin-A; however, corpus lutea present on proestrus and estrus bound little or no [125I]rh-activin-A. [125I]rh-Activin-A-binding sites were also present in the uterus and oviduct in a cycle-dependent manner. The highest levels of binding were found in the muscle wall of the uterus and the epithelial lining of the thick-walled portion of the uterus on metestrus and diestrus. In addition, [125I]rh-activin-A bound to the cumulus-oocyte complex present in the oviduct on metestrus, but did not bind to the oocytes present in developing follicles. Binding of [125I]rh-inhibin-A was restricted to the antral granulosa cells of 450- to 500-microns follicles. No other ovarian, uterine, or oviduct cells bound [125I]rh-inhibin-A. [125I]rh-Activin-A and [125I]rh-inhibin-A ligand binding was associated primarily with follicles coexpressing inhibin/activin subunit and follistatin mRNA.(ABSTRACT TRUNCATED AT 400 WORDS)

Morphometric study of ovarian follicular growth during the estrous cycle of the laboratory-maintained Indian desert gerbil, Meriones hurrianae (Jerdon).

The pattern of changes in the population of healthy and regressing follicles in relation to the estrous cycle, was studied in the recently domesticated desert gerbil, Meriones hurrianae, which exhibits a longer cycle (proestrus: 1.5 days; estrus: 1 day; metestrus: 1.5 days; diestrus: 3 days). As the granulosa cell layers were not distinct, the follicles were classified on the basis of their morphology and size (group I: small preantral follicles; group II: large preantral follicles; group III: small antral follicles; group IV: medium-sized antral follicles; group V: large-sized follicles; group VI: Graafian follicles). Regressing follicles were classified likewise. Five animals were used for each stage of the cycle. One ovary from each gerbil was serially cut and stained with hematoxylin-eosin. Healthy and regressing follicles were found throughout the cycle. The follicular development occurred typically in a biphasic manner. In phase A, in which follicles reach the antral stage, the growth of oocytes and follicles is linearly correlated, whereas, in phase B, oocyte size remains stationary while follicle size increases due to accumulation of antral fluid. However, oocyte growth and nucleus growth are positively and linearly correlated (r = 0.74). The healthy follicles under 260 microns (groups I, II) do not fluctuate in the ovary of cyclic gerbils, whereas follicles in group III (261-350 microns) increase significantly (p < 0.05) at estrus and those of group IV in diestrus. Further, healthy group III follicles are reduced in diestrus while regressing follicles in group III increase. These findings thus suggest that the antral follicles are rescued from atresia at estrus.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of the Neural Cell Adhesion Molecule in Endocrine Cells of the Ovary*

In the adult mammalian ovary morphogenesis and differentiation processes are under hormonal control and, thus, occur in a highly regulated way during the sexual cycle. Cell-cell interactions, such as cell adhesion and cell separation, are crucial during these events. Here we show that the ovarian endocrine cells, which are prototypes of steroid-producing cells, express neural cell adhesion molecules (NCAMs). The combined use of in situ hybridization histochemistry, immunocytochemistry at the light and electron microscope levels, S1 nuclease protection assays, and Western blotting revealed that in the ovary of the adult rat during the estrus cycle and pregnancy, NCAM mRNA and the 140-kDa isoform of this protein are expressed mainly in granulosa cells of growing preantral and antral follicles and in corpora lutea. Since the granulosa cells lining the forming antrum and the antral fluid were strongly immunoreactive, a role for NCAM in the formation of the follicular antrum is proposed. The expression of NCAM was also associated with luteal cells of the active corpus luteum, indicating a role for NCAM in the morphogenesis of this endocrine compartment. Moreover, thecal cells of large follicles and hypertrophic thecal cells of atretic follicles expressed NCAM, as did interstitial cells, which are derived from thecal cells of atretic follicles. We propose that the adhesion molecule, NCAM, is an important factor involved in the recognition and intercellular interaction of ovarian endocrine cells and, thus, participates in the regulation of the cyclic remodeling processes of the ovarian endocrine compartments.

Concentrations of follicle regulatory protein, steroids and gonadotrophins in antral fluids from women stimulated with metrodin and hCG

Concentrations of immunoreactive follicle regulatory protein (FRP) were determined in 184 follicular fluid samples recovered from 30 patients in whom ovarian stimulation before oocyte recovery and in-vitro fertilization was induced with FSH (150 i.u./day). Ovum recovery was scheduled when the diameters of greater than or equal to 2 follicles reached 15-17 mm and serum oestradiol values were 740 pM/follicle. The mean level of FRP in fluid from follicles yielding oocytes which fertilized and cleaved within 48 h of recovery (24.4 +/- 3.08 immunoreactive units [IRU]/ml; 1 IRU = approximately 1 ng pig FRP) was higher than that in fluid from follicles yielding oocytes which did not fertilize (10.5 +/- 1.67 IRU/ml, P less than 0.05). FRP in fluid from follicles yielding oocytes which fertilized but did not cleave within 48 h of recovery was 17.2 +/- 2.89 IRU/ml. Overall, concentrations of FRP did not correlate with follicular fluid volume or with FSH or LH concentrations, but were positively related to prolactin, oestradiol and total protein levels (P less than 0.04). The results indicate that the FRP content of follicular fluid may be predictive of follicle/oocyte maturity. A dose-dependent increase in release of FRP by pig granulosa cells cultured in medium supplemented with 10-100 ng prolactin/ml was demonstrated. Prolactin may, therefore, be an important determinant of FRP production by granulosa cells.